Content Status
Type
Linked Node
Concept of Polymerase Chain Reaction [PCR] Amplification of the Target
Learning ObjectivesPCR
H5Content
Content
GeneXpert MTB/RIF assay targets rpoB gene that encodes for RNA polymerase β subunit.
- More than 95% of mutations in the rpoB gene are located in the 81 base pair core regions (codons 507 – 533) also known as Rifampicin Resistance Determining Region (RRDR).
- Mutations lead to inhibition of binding of rifampicin to RNA polymerase. This results in the development of RIF resistance by the mycobacteria.
- Semi-nested polymerase chain reaction (PCR) using a set of three primers P1, P2 and P3 for amplification of 192 bp of rpoB gene containing 81 bp RRDR
- 1st round of amplification takes place using primers P1 and P3 (19 cycles)
- In the next round, P2 and P3 are used to amplify the target (40 cycles)
Resources
Kindly provide your valuable feedback on the page to the link provided HERE
LMS Page Link
Content Creator
Reviewer
- Log in to post comments